Loading history changes the morphology and compressive force-induced expression of receptor activator of nuclear factor kappa B ligand/osteoprotegerin in MLO-Y4 osteocytes.
Loading history changes the morphology and compressive force-induced expression of receptor activator of nuclear factor kappa B ligand/osteoprotegerin in MLO-Y4 osteocytes.
复制标题
作者：
Wang Z;Weng Y;Ishihara Y;Odagaki N;Ei Hsu Hlaing E;Izawa T;Okamura H;Kamioka H
在这项研究中，我们研究了机械负荷历史对核因子κ B配体受体激活因子（RANKL）和骨保护素（OPG）在MLO-Y 4骨细胞样细胞中表达的影响。接种MLO-Y 4骨细胞后3小时，在骨细胞上施加31达因/cm 2的连续压缩力（CCF），有或没有额外的CCF（32达因/cm 2）。36 h后，移除额外CCF（加载历史），恢复期为10 h。在0、3、6和10 h检测RANKL、OPG的表达、RANKL/OPG比值、细胞数量、存活率和形态学。然后，将相同的另外的CCF再次应用于所有骨细胞，持续1小时，其中有或没有差距连接抑制剂，以检查RANKL、OPG、RANKL/OPG比率和表征MLO-Y 4细胞表型所必需的其他基因的表达。采用光漂白后荧光恢复技术检测MLO-Y 4细胞间缝隙连接通讯（GJIC）的差异。RANKL和OPG的表达的MLO-Y 4骨细胞无负荷的历史显着减少和增加，分别响应于1小时的额外重量的负荷。然而，在有负荷史但没有间隙连接抑制剂处理的MLO-Y 4骨细胞中，RANKL、OPG的表达和RANKL/OPG的比值维持在与对照组相同的水平。负荷史处理显著改变GJIC的能力和连接蛋白43（Cx43）的蛋白表达，但不影响Cx43的mRNA表达。在有和没有加载历史的MLO-Y 4骨细胞样细胞之间或在恢复期的不同时间检查点之间，未观察到细胞数量或活力的显著差异。恢复期细胞形态发生明显变化，并与OPG、Gja 1和Dmp 1的表达相关。我们的研究结果表明，压力诱导的RANKL/OPG表达的变化可以在至少11小时内习惯于36小时的CCF暴露。GJIC和细胞形态可能在MLO-Y 4骨细胞样细胞对负荷历史的反应中起作用。
In this study, we investigated the effect of the mechanical loading history on the expression of receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) in MLO-Y4 osteocyte-like cells. Three hours after MLO-Y4 osteocytes were seeded, a continuous compressive force (CCF) of 31 dynes/cm2 with or without additional CCF (32 dynes/cm2) was loaded onto the osteocytes. After 36 h, the additional CCF (loading history) was removed for a recovery period of 10 h. The expression of RANKL, OPG, RANKL/OPG ratio, cell numbers, viability and morphology were time-dependently examined at 0, 3, 6 and 10 h. Then, the same additional CCF was applied again for 1 h to all osteocytes with or without the gap junction inhibitor to examine the expression of RANKL, OPG, the RANKL/OPG ratio and other genes that essential to characterize the phenotype of MLO-Y4 cells. Fluorescence recovery after photobleaching technique was also applied to test the differences of gap-junctional intercellular communications (GJIC) among MLO-Y4 cells. The expression of RANKL and OPG by MLO-Y4 osteocytes without a loading history was dramatically decreased and increased, respectively, in response to the 1-h loading of additional weight. However, the expression of RANKL, OPG and the RANKL/OPG ratio were maintained at the same level as in the control group in the MLO-Y4 osteocytes with a loading history but without gap junction inhibitor treatment. Treatment of loading history significantly changed the capacity of GJIC and protein expression of connexin 43 (Cx43) but not the mRNA expression of Cx43. No significant difference was observed in the cell number or viability between the MLO-Y4 osteocyte-like cells with and without a loading history or among different time checkpoints during the recovery period. The cell morphology showed significant changes and was correlated with the expression of OPG, Gja1 and Dmp1 during the recovery period. Our findings indicated that the compressive force-induced changes in the RANKL/OPG expression could be habituated within at least 11 h by 36-h CCF exposure. GJIC and cell morphology may play roles in response to loading history in MLO-Y4 osteocyte-like cells.
登录
查看更多内容
影响因子：
4
作者：
McDiarmid, Troy A.;Yu, Alexj;Rankin, Catharine H.
通讯作者：
Rankin, Catharine H.
影响因子：
6.2
作者：
Bivi, Nicoletta;Condon, Keith W.;Allen, Matthew R.;Farlow, Nathan;Passeri, Giovanni;Brun, Lucas R.;Rhee, Yumie;Bellido, Teresita;Plotkin, Lilian I.
通讯作者：
Plotkin, Lilian I.
影响因子：
82.9
作者：
Nakashima, Tomoki;Hayashi, Mikihito;Takayanagi, Hiroshi
通讯作者：
Takayanagi, Hiroshi
影响因子：
15.9
作者：
Lotinun, Sutada;Ishihara, Yoshihito;Baron, Roland
通讯作者：
Baron, Roland
影响因子：
2.8
作者：
Bivi, Nicoletta;Pacheco-Costa, Rafael;Brun, Lucas R.;Murphy, Thomas R.;Farlow, Nathan R.;Robling, Alexander G.;Bellido, Teresita;Plotkin, Lilian I.
通讯作者：
Plotkin, Lilian I.